Phosphoproteomic profiling of human SH-SY5Y neuroblastoma cells during response to 6-hydroxydopamine-induced oxidative stress
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- 作者：Megumi Nakamura ; Masaki Yamada ; Takako Ohsawa ; Hiraku Morisawa ; Tsutomu Nishine ; Osamu Nishimura and Tosifusa Toda
- 关键词：Oxidative stress ; SH-SY5Y cells ; Phosphoproteome ; 6-hydroxydopamine
- 刊名：Biochimica et Biophysica Acta (BBA)/Molecular Cell Research
- 出版年：2006
- 出版时间：September 2006
- 年：2006
- 卷：1763
- 期：9
- 页码：977-989
- 全文大小：792 K
文摘
Dopaminergic neurons are known to be vulnerable to age-related neuronal disorders due to reactive oxygen species (ROS) generated during dopamine metabolism. However, it remains unclear what kinds of proteins are involved in the response to oxidative stress. We examined changes in whole proteins and phosphoproteins in the human dopaminergic neuroblastoma cell line SH-SY5Y under oxidative stress induced by the dopaminergic neurotoxin 6-hydroxydopamine (6-OHDA). Proteins of SH-SY5Y cells at various stages of oxidative stress were separated by two-dimensional gel electrophoresis for comparative analysis. Increase in glutathione-S-transferase pi was detected on SYPRO Ruby-stained gels by computer-aided image analysis. Stress-induced alterations in phosphoproteins were detected by Pro-Q Diamond staining. Elongation factor 2, lamin A/C, T-complex protein 1, and heterogeneous nuclear ribonucleoprotein H3 were identified by MALDI-TOF mass spectrometry as stress-responsive elements.